Part:BBa_J45014:Design

alcohol acetyltransferase I; converts isoamyl alcohol to isoamyl acetate (banana odor)

Assembly Compatibility:

• 10
  COMPATIBLE WITH RFC[10]
• 12
  COMPATIBLE WITH RFC[12]
• 21
  INCOMPATIBLE WITH RFC[21]

  Illegal BglII site found at 99
  Illegal BamHI site found at 1426
  
• 23
  COMPATIBLE WITH RFC[23]
• 25
  COMPATIBLE WITH RFC[25]
• 1000
  INCOMPATIBLE WITH RFC[1000]

  Illegal SapI.rc site found at 526
  Illegal SapI.rc site found at 1534
  

Design Notes

• Constructed by PCR.
• Eliminated EcoRI site at base pair 412 by changing base pair 414 from A to G
• BBa_J45014 differs from BBa_J45006 by a single point mutation at base pair 1148 (T -> C).

Template

Saccharomyces cerevisiae strain ACLY387 genome

Forward primer

5'- GTT TCT TCG AAT TCG CGG CCG CTT CTA GAT GAA TGA AAT CGA TGA GAA AAA TC -3'

Reverse primer

5'- GTT TCT TCC TGC AGC GGC CGC TAC TAG TAT TAT TAa ggg cct aaa agg aga gct ttg -3'

Forward mutagenesis primer

5'- GAA GCA AAT ATT AGA AGA GTT CAA AAA TAG TAA GGG -3'

Reverse mutagenesis primer

5'- CCC TTA CTA TTT TTG AAC TCT TCT AAT ATT TGC TTC -3'

Source

ATF1 gene from Saccharomyces cerevisiae (GenBank [http://www.ncbi.nlm.nih.gov/entrez/viewer.fcgi?db=Nucleotide&dopt=GenBank&val=1420813 Z75285])

References

<biblio>

1. Fujii--1984 pmid=8085822
2. Horton03 pmid=12937998
3. Singh--2007 pmid=17891501

</biblio>